Galactokinase inhibitors for the treatment and prevention of associated diseases and disorders

ABSTRACT

Disclosed are inhibitors of human galactokinase of formula (1) that are useful in treating or preventing a galactokinase mediated disease or disorder, e.g., galactosemia. Also disclosed are a composition comprising a pharmaceutically acceptable carrier and at least one inhibitor of the invention, and a method of treating or preventing such disease or disorder in a mammal. Formula (I).

CROSS-REFERENCE TO A RELATED APPLICATION

This patent application is a U.S. National Phase of International PatentApplication No. PCT/US11/53021, filed Sep. 23, 2011, which isincorporated by reference.

BACKGROUND OF THE INVENTION

Galactose is an abundant hexose existing as lactose in milk, dairyproducts, fruits, vegetables and many other foods (Acosta, P. B. and K.C. Gross, Eur. J. Pediatr, 1995. 154(7 Suppl. 2): p. S87-92; Berry, G.T., et al., J. Inherit. Metab. Dis, 1993. 16(1): p. 91-100). It ismetabolized through an evolutionarily conserved pathway referred to asthe Leloir pathway (Leloir, L. F., Arch Biochem, 1951. 33(2): p.186-90). The first enzyme of the pathway, galactokinase (GALK), convertsα-D-galactose to galactose-1-phosphate (gal-1-p) (Atkinson, M. R., E.Johnson, and R. K. Morton, Nature, 1959. 184: p. 1925-7). Then, in thepresence of the second enzyme, galactose-1-phosphate uridyltransferase(GALT), gal-1-P will react with UDP-glucose to form UDP-galactose andglucose-1-phosphate (Arabshahi, A., et al., Biochemistry, 1986. 25(19):p. 5583-9).

Deficiency of GALT results in a potentially lethal disorder calledClassic Galactosemia (CG) (Isselbacher, K. J., et al., Science, 1956.123(3198): p. 635-6; Rennert, O. M., Annals of clinical and laboratoryscience, 1977. 7(6): p. 443-8). Patients with CG accumulate high levelof gal-1-p which can result in severe disease during the newborn period,including liver failure, coagulopathy, coma, and death if not treated(Goppert, F., Klin. Wschr., 1917(54): p. 473-477; Mason H, T. M., Am. J.Dis. Child., 1935(50): p. 359-374; Tyfield, L., et al., Hum. Mutat.,1999. 13(6): p. 417-30). Although removal of galactose from the diet canprevent neonatal death, CG patients still develop chronic complicationssuch as premature ovarian insufficiency (POI), ataxia, speech dyspraxiaand mental retardation even in galactose-restricted diet (Waggoner, D.,Buist, N R M, Donnell, G N, Journal of Inherited Metabolic Disorders,1990. 13: p. 802-818; Waggoner, D., Buist, N R M, InternationalPediatrics, 1993. 8: p. 97-100).

The mechanisms for the above chronic complications remain uncertain, butseveral lines of evidences indicate accumulation of gal-1-p is a majorfactor that contributes to these complications. Except for cataracts,patients with an inherited deficiency of GALK do not experience thecomplications observed in GALT-deficient patients (Gitzelmann, R., H. J.Wells, and S. Segal, Eur. J. Clin. Invest., 1974. 4(2): p. 79-84;Gitzelmann, R., J. Pediatr., 1975. 87(6 Pt 1): p. 1007-8). Similarly,while gal7 (i.e., GALT-deficient) mutant yeast stops growing upongalactose challenge, a gal 7 gal1 double mutant strain (i.e., GALT- andGALK-deficient) is no longer sensitive to galactose (Douglas, H. C. andD. C. Hawthorne, Genetics, 1964. 49: p. 837-44; Douglas, H. C. and D. C.Hawthorne, Genetics, 1966. 54(3): p. 911-6). A significant amount ofgalactose is found in non-dairy foods such as vegetables and fruits, andmore importantly, galactose is also produced endogenously from thenatural turnover of glycolipids and glycoproteins. Moreover, usingisotope labeling, Berry et al. demonstrated that a 50 kg adult malecould produce up to 2 grams of galactose per day (Berry, G. T., et al.,Mol. Genet. Metab., 2004. 81(1): p. 22-30; Berry, G. T., et al., Eur. J.Pediatr., 1997. 156 Suppl. 1: p. S43-9; Berry, G. T., et al., Lancet,1995. 346(8982): p. 1073-4).

In addition, it is known that GALKmodifies the PTEN/AKT pathway in anumber of human tissues and human cell lines. The galactose-1-phosphateproduced by GALK feeds into glycolysis. GALK1 is over-expressed in anumber of tumors. Thus, inhibition of GALK may down regulate thePTEN/ATK pathway and therefore interfere with tumor growth and/ordevelopment.

The foregoing shows that there is an unmet need for inhibitors ofgalactokinase enzyme and agents for the prophylaxis and/or therapy ofdiseases preventable or treatable by inhibiting the enzyme.

BRIEF SUMMARY OF THE INVENTION

The invention provides galactokinase inhibitors, e.g., a compound offormula (I):

wherein R¹ and R² are each independently selected from the groupconsisting of hydrogen, alkyl, aryl, and heteroaryl or wherein R¹ andR², taken together along with the carbon atoms to which they areattached, form a 5- to 7-membered carbocyclic or heterocyclic ring,

wherein R³ is selected from the group consisting of —NH-alkyl,—NH-cycloalkyl, —NH-aryl, —NH-alkylaryl, —NH-heteroaryl, —N-heteroaryl,and —NR¹²R¹³ wherein R¹² and R¹³ are each independently selected fromthe group consisting of hydrogen, alkyl, cycloalkyl, aryl, arylalkyl,and heteroaryl, or wherein R¹² and R¹³ together form a heteroaryl or aheterocycloalkyl,

wherein R⁴ is selected from the group consisting of hydrogen and alkyl,or

wherein R³ and R⁴ together form a group of the formula:

wherein R⁶, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ are each independently selectedfrom the group consisting of hydrogen, alkyl, aryl, and heteroaryl,

wherein m is 0 or 1,

wherein R⁵ is heteroaryl,

wherein alkyl, aryl, heterocycloalkyl, carbocyclic ring, heterocyclicring, arylalkyl, and heteroaryl groups are unsubstituted or optionallysubstituted with one or more substituents selected from the groupconsisting of alkyl, aryl, halo, trifluoromethyl, alkoxy, aryloxy,amino, alkylamino, and dialkylamino,

or a pharmaceutically acceptable salt thereof.

The invention also provides a pharmaceutical composition comprising acompound or salt of the invention and a pharmaceutically acceptablecarrier.

The invention further provides method for treating or preventing a humangalactokinase mediated disease or disorder in a mammal, comprisingadministering to the mammal in need thereof, a therapeutically orprophylactically effective amount of a compound of the invention or saltthereof.

The invention additionally provides a method for treating or preventinga PTEN/AKT mis-regulated cancer in a mammal, comprising administering tothe mammal in need of, a therapeutically or prophylactically effectiveamount of a compound of the invention, salt thereof, enantiomersthereof, a mixture of enantiomers thereof, or diastereomers thereof of acompound.

The invention further provides a method of reducing the level ofgalactose-1-phosphate in a cell, comprising contacting the cell witheffective amount of a compound of the invention, salt thereof,enantiomers thereof, a mixture of enantiomers thereof, or diastereomersthereof.

DETAILED DESCRIPTION OF THE INVENTION

In accordance with an embodiment, the invention provides a compound offormula (I):

wherein R¹ and R² are each independently selected from the groupconsisting of hydrogen, alkyl, aryl, and heretoaryl or wherein R¹ andR², taken together along with the carbon atoms to which they areattached, form a 5- to 7-membered carbocyclic or heterocyclic ring,

wherein R³ is selected from the group consisting of —NH-alkyl,—NH-cycloalkyl, —NH-aryl, —NH-alkylaryl, —NH-heteroaryl, —N-heteroaryl,and —NR¹²R¹³ wherein R¹² and R¹³ are each independently selected fromthe group consisting of hydrogen, alkyl, cycloalkyl, aryl, arylalkyl,and heteroaryl, or wherein R¹² and R¹³ together form a heteroaryl or aheterocycloalkyl,

wherein R⁴ is selected from the group consisting of hydrogen and alkyl,or

wherein R³ and R⁴ together form a group of the formula:

wherein R⁶, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ are each independently selectedfrom the group consisting of hydrogen, alkyl, aryl, and heteroaryl,

wherein m is 0 or 1,

wherein R⁵ is heteroaryl,

wherein alkyl, aryl, heterocycloalkyl, carbocyclic ring, heterocyclicring, arylalkyl, and heteroaryl groups are unsubstituted or optionallysubstituted with one or more substituents selected from the groupconsisting of alkyl, aryl, halo, trifluoromethyl, alkoxy, aryloxy,amino, alkylamino, and dialkylamino,

with the proviso that when R⁵ is benzoxazol-2-yl, R¹ is hydrogen, R² isphenyl, R³ and R⁴ together form

and m is 1, R¹⁰ and R¹¹ are not simultaneously methyl,

or a pharmaceutically acceptable salt thereof.

Referring now to terminology used generically herein, the term “alkyl”means a straight-chain or branched alkyl substituent containing from,for example, 1 to about 6 carbon atoms, preferably from 1 to about 4carbon atoms, more preferably from 1 to 2 carbon atoms. Examples of suchsubstituents include methyl, ethyl, propyl, isopropyl, n-butyl,sec-butyl, isobutyl, tert-butyl, pentyl, isoamyl, hexyl, and the like.

The term “cycloalkyl,” as used herein, means a cyclic alkyl substituentcontaining from, for example, about 3 to about 8 carbon atoms,preferably from about 43 to about 7 carbon atoms, and more preferablyfrom about 5 to about 6 carbon atoms. Examples of such substituentsinclude cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,cyclooctyl, and the like. The cyclic alkyl groups may be unsubstitutedor further substituted with one or more alkyl groups such as methylgroups, ethyl groups, and the like.

The term “heteroaryl,” as used herein, refers to a 5, 6, or 7-memberedaromatic ring system containing one or more heteroatoms selected fromthe group consisting of O, N, S, and combinations thereof. Theheteroaryl group can be any suitable heteroaryl group. The heteroarylgroup can be a monocyclic heteroaryl group or a bicyclic heteroarylgroup. Suitable bicyclic heteroaryl groups include monocylic heteroarylrings fused to a C₆-C₁₀ aryl ring. It is understood that a 6-memberedheteroaryl group comprises 4n+2π electrons, according to Hückel's Rule,and that a 5-, 7-, and 8-membered heteroaryl group has six electronsprovided from a combination of p orbitals and an unshared pair ofelectrons provided by a heteroatom or heteroatoms which occupy bondingorbitals and constitute an aromatic sextet. Non-limiting examples ofsuitable heteroaryl groups include furanyl, thiophenyl, pyrrolyl,pyrazolyl, imidazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, isoxazolyl,oxazolyl, isothiazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl,triazinyl, benzofuranyl, benzothiophenyl, indolyl, quinolinyl,isoquinolinyl, benzimidazolyl, benzoxazolinyl, benzothiazolinyl, andquinazolinyl. The heteroaryl group can be linked at any open position ofthe heteroaryl group. For example, the furanyl group can be a furan-2-ylgroup or a furan-3-yl group, and the thiophenyl group can be athiophene-2-yl group or a thiophene-3-yl group. The heteroaryl group isoptionally substituted with 1, 2, 3, 4, or 5 substituents as recitedherein, wherein the optional substituent can be present at any openposition on the heterocyclyl group.

The term “halo” or “halogen,” as used herein, means a substituentselected from Group VIIA, such as, for example, fluorine, bromine,chlorine, and iodine.

The term “aryl” refers to an unsubstituted or substituted aromaticcarbocyclic substituent, as commonly understood in the art, and the term“C₆-C₁₀ aryl” includes phenyl and naphthyl. It is understood that theterm aryl applies to cyclic substituents that are planar and comprise4n+2π electrons, according to Hückel's Rule.

The term “alkylaryl” refers to an unsubstituted or substituted alkylgroup bonded to an unsubstituted or substituted aryl group. Thealkylaryl group is typically bonded to the core structure of themolecule by way of the alkyl portion of the alkylaryl group.

The term “heterocycloalkyl,” as used herein, means a non-aromatic cyclicalkyl substituent containing a heteroatom and further containing from,for example, about 3 to about 7 carbon atoms, preferably from about 3 toabout 6 carbon atoms, and more preferably from about 3 to about 5 carbonatoms. The heterocycloalkyl group can be monocyclic or can be fused toanother ring, wherein the other ring can be a cycloalkyl ring, an arylring, or another heterocycloalkyl ring. Examples of such substituentsinclude tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl,piperidinyl, tetrahydroazepinyl, and the like. The heterocycloalkylgroups may be unsubstituted or further substituted with alkyl groupssuch as methyl groups, ethyl groups, and the like.

In certain embodiments of the compound of formula (I), R⁵ is selectedfrom the group consisting of benzoxazol-2-yl, 5-bromo-benzoxazol-2-yl,5-methyl-benzoxazol-2-yl, 6-methyl-benzoxazol-2-yl,6-phenyl-benzoxazol-2-yl, benzoimidazol-2-yl, benzothiazol-2-yl,indol-1-yl, indol-2-yl, indol-3-yl, furan-2-yl, furan-3-yl,thiophene-2-yl, thiophene-3-yl, imidazol-1-yl, imidazol-4-yl,thiazol-2-yl, thiazol-4-yl, pyrazol-3-yl, pyrazol-4-yl, pyridin-2-yl,pyridin-3-yl, and pyridin-4-yl.

In preferred embodiments, R⁵ is selected from the group consisting ofbenzoxazol-2-yl, 5-bromo-benzoxazol-2-yl, 5-methyl-benzoxazol-2-yl,6-methyl-benzoxazol-2-yl, and 6-phenyl-benzoxazol-2-yl.

In a more preferred embodiment, R⁵ is benzoxazol-2-yl.

In accordance with an embodiment, R³ and R⁴ together form

wherein m is 1, and wherein the compound has the formula (Ia):

In accordance with another embodiment of the compound of formula (Ia),R¹ is hydrogen and R² is phenyl optionally substituted with one or moresubstituents selected from the group consisting of halogen,trifluoromethyl, alkyl, alkoxy, aryloxy, and dialkylamino.

In accordance with another embodiment of the compound of formula (Ia),R¹ is hydrogen and R² is heteroaryl optionally substituted with one ormore substituents selected from the group consisting of halogen,trifluoromethyl, alkyl, alkoxy, aryloxy, and dialkylamino.

In accordance with an embodiment, specific examples of the formula (Ia)include those wherein R⁶, R⁷, R¹⁰, and R¹¹ are hydrogen.

In accordance with another embodiment, specific examples of the formula(Ia) include those wherein R⁸ and R⁹ are both hydrogen.

In accordance with another embodiment, specific examples of the formula(Ia) include those wherein R⁸ and R⁹ are both methyl.

In accordance with another embodiment, specific examples of the formula(Ia) include those wherein R⁸ is hydrogen and R⁹ is hydrogen or phenyloptionally substituted with one or more substituents selected from thegroup consisting of halo, trifluoromethyl, alkyl, alkoxy, aryloxy,amino, alkylamino, and dialkylamino, or heteroaryl optionallysubstituted with one or more substituents selected from the groupconsisting of halo, trifluoromethyl, alkyl, alkoxy, aryloxy, amino,alkylamino, and dialkylamino.

In accordance with preferred embodiments, specific examples of theformula (Ia) include those wherein R⁹ is phenyl substituted with one ormore substituents selected from the group consisting of halo,trifluoromethyl, alkyl, alkoxy, and dialkylamino.

In accordance with other preferred embodiments, specific examples of theformula (Ia) include those wherein R⁹ is heteroaryl selected from thegroup consisting of 5-methylthiophen-2-yl, pyridine-3-yl, pyridine-4-yl,2-chloropyridin-4-yl, 3-trifluoropyridin-2-yl,4-trifluoromethylpyridin-3-yl, 2-chloropyridin-3-yl,2-bromopyridin-3-yl, 3-methylthophen-2-yl, 3-bromopyridin-4-yl,4-bromopyrazol-3-yl, 4-bromo-1-methylpyrazol-3-yl, 3-bromopyridin-4-yl,4-chloro-1-methylpyrazol-3-yl, pyrazol-3-yl, 5-methylpyrazol-3-yl, and4-chloro-1-methylpyrazol-3-yl.

In keeping with the embodiments described above, specific examples ofcompounds include compounds selected from the group consisting of:

In accordance with an embodiment, the compound of formula (I) has theformula (Ib):

wherein X is selected from the group consisting of O, S, and SO₂,wherein R¹⁴ is selected from the group consisting of hydrogen, alkyl,and aryl, and wherein n is 0, 1, or 2.

In accordance with an embodiment of the compound of formula (Ib), m is1.

In an embodiment of these compounds, R⁶, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ arehydrogen.

In a preferred embodiment of these compounds, R⁸ and R⁹ are bothhydrogen.

In another preferred embodiment of these compounds, R⁸ and R⁹ are bothmethyl.

In another preferred embodiment of these compounds, R⁸ is hydrogen andR⁹ is phenyl.

In keeping with the embodiments described above, specific examples ofcompounds include compounds selected from the group consisting of:

In accordance with an embodiment of the compound of formula (Ib), m is0.

In an embodiment of these compounds, R⁶, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ arehydrogen.

In accordance with a specific embodiment of these compounds, thecompound is

In accordance with an embodiment of the compounds of formula (I), R³ isselected from the group consisting of —NH-alkyl, —NH-cycloalkyl,—NH-aryl, —NH-alkylaryl, —NH-heteroaryl, —N-heteroaryl, and —NR¹²R¹³wherein R¹² and R¹³ are each independently selected from the groupconsisting of hydrogen, alkyl, cycloalkyl, aryl, arylalkyl, andheteroaryl, or wherein R¹² and R¹³ together form a heteroaryl or aheterocycloalkyl,

wherein R⁴ is selected from the group consisting of hydrogen and alkyl,wherein R⁵ is heteroaryl, and wherein alkyl, aryl, heterocycloalkyl,carbocyclic ring, heterocyclic ring, arylalkyl, and heteroaryl groupsare unsubstituted or optionally substituted with one or moresubstituents selected from the group consisting of alkyl, aryl, halo,trifluoromethyl, alkoxy, aryloxy, amino, alkylamino, and dialkylamino.

In keeping with the embodiments described above, specific examples ofcompounds include compounds selected from the group consisting of:

In accordance with an embodiment, the compound of formula (I) has theformula (Ic):

wherein R³ is selected from the group consisting of —NH-alkyl,—NH-cycloalkyl, —NH-aryl, —NH-alkylaryl, —NH-heteroaryl, —N-heteroaryl,and —NR¹²R¹³ wherein R¹² and R¹³ are each independently selected fromthe group consisting of hydrogen, alkyl, cycloalkyl, aryl, arylalkyl,and heteroaryl, or wherein R¹² and R¹³ together form a heteroaryl orheterocycloalkyl,

wherein R⁴ is selected from the group consisting of hydrogen and alkyl,

wherein Y is selected from the group consisting of CHR¹⁰, O, S, and SO₂,

wherein R¹⁰ and R¹⁴ are each independently selected from the groupconsisting of hydrogen, alkyl, and aryl optionally substituted with oneor more substituents selected from the group consisting of halo,trifluoromethyl, alkyl, alkoxy, aryloxy, and dialkylamino, and

wherein o is 0, 1, or 2.

In keeping with the embodiments described above, specific examples ofcompounds include compounds selected from the group consisting of:

It will be understood that the position of double bonds in adihydropyrimidinyl ring is not fixed and that any structure comprising adihydropyrimidinyl ring also includes tautomers thereof. For example,the structure

also is intended to refer to the tautomers

when R¹ and R² are not hydrogen;

and also when R¹ and/or R² are hydrogen to the tautomer

In any of the above embodiments, the compound or salt of formula (I) canhave at least one asymmetric carbon atom. When the compound or salt hasat least one asymmetric carbon atom, the compound or salt can exist inthe racemic form, in the form of its pure optical isomers, or in theform of a mixture wherein one isomer is enriched relative to the other.In particular, in accordance with the present invention, when theinventive compounds have a single asymmetric carbon atom, the inventivecompounds may exist as racemates, i.e., as mixtures of equal amounts ofoptical isomers, i.e., equal amounts of two enantiomers, or in the formof a single enantiomer. As used herein, “single enantiomer” is intendedto include a compound that comprises more than 50% of a singleenantiomer (i.e., enantiomeric excess up to 100% pure enantiomer).

When the compound or salt has more than one chiral center, the compoundor salt can therefore exist as a mixture of diastereomers or in the formof a single diastereomer. As used herein, “single diastereomer” isintended to mean a compound that comprises more than 50% of a singlediastereomer (i.e., diastereomeric excess to 100% pure diastereomer).

The phrase “pharmaceutically acceptable salt” is intended to includenontoxic salts synthesized from the parent compound which contains abasic or acidic moiety by conventional chemical methods. Generally, suchsalts can be prepared by reacting the free acid or base forms of thesecompounds with a stoichiometric amount of the appropriate base or acidin water or in an organic solvent, or in a mixture of the two.Generally, nonaqueous media such as ether, ethyl acetate, ethanol,isopropanol, or acetonitrile are preferred. Lists of suitable salts arefound in Remington's Pharmaceutical Sciences, 18th ed., Mack PublishingCompany, Easton, Pa., 1990, p. 1445, and Journal of PharmaceuticalScience, 66, 2-19 (1977).

Suitable bases include inorganic bases such as alkali and alkaline earthmetal bases, e.g., those containing metallic cations such as sodium,potassium, magnesium, calcium and the like. Non-limiting examples ofsuitable bases include sodium hydroxide, potassium hydroxide, sodiumcarbonate, and potassium carbonate. Suitable acids include inorganicacids such as hydrochloric acid, hydrobromic acid, hydroiodic acid,sulfuric acid, phosphoric acid, and the like, and organic acids such asp-toluenesulfonic, methanesulfonic acid, benzenesulfonic acid, oxalicacid, p-bromophenylsulfonic acid, carbonic acid, succinic acid, citricacid, benzoic acid, acetic acid, maleic acid, tartaric acid, fattyacids, long chain fatty acids, and the like. Preferred pharmaceuticallyacceptable salts of inventive compounds having an acidic moiety includesodium and potassium salts. Preferred pharmaceutically acceptable saltsof inventive compounds having a basic moiety (e.g., a quinoline group ora dimethylaminoalkyl group) include hydrochloride and hydrobromidesalts. The compounds of the present invention containing an acidic orbasic moiety are useful in the form of the free base or acid or in theform of a pharmaceutically acceptable salt thereof.

It should be recognized that the particular counterion forming a part ofany salt of this invention is usually not of a critical nature, so longas the salt as a whole is pharmacologically acceptable and as long asthe counterion does not contribute undesired qualities to the salt as awhole.

It is further understood that the above compounds and salts may formsolvates, or exist in a substantially uncomplexed form, such as theanhydrous form. As used herein, the term “solvate” refers to a molecularcomplex wherein the solvent molecule, such as the crystallizing solvent,is incorporated into the crystal lattice. When the solvent incorporatedin the solvate is water, the molecular complex is called a hydrate.Pharmaceutically acceptable solvates include hydrates, alcoholates suchas methanolates and ethanolates, acetonitrilates and the like. Thesecompounds can also exist in polymorphic forms.

The present invention is further directed to a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and atleast one compound or salt described herein.

It is preferred that the pharmaceutically acceptable carrier be one thatis chemically inert to the active compounds and one that has nodetrimental side effects or toxicity under the conditions of use.

The choice of carrier will be determined in part by the particularcompound of the present invention chosen, as well as by the particularmethod used to administer the composition. Accordingly, there is a widevariety of suitable formulations of the pharmaceutical composition ofthe present invention. The following formulations for oral, aerosol,nasal, pulmonary, parenteral, subcutaneous, intravenous, intraarterial,intramuscular, intraperitoneal, intrathecal, intratumoral, topical,rectal, and vaginal administration are merely exemplary and are in noway limiting.

The pharmaceutical composition can be administered parenterally, e.g.,intravenously, intraarterially, subcutaneously, intradermally, orintramuscularly. Thus, the invention provides compositions forparenteral administration that comprise a solution or suspension of theinventive compound or salt dissolved or suspended in an acceptablecarrier suitable for parenteral administration, including aqueous andnon-aqueous isotonic sterile injection solutions.

Overall, the requirements for effective pharmaceutical carriers forparenteral compositions are well known to those of ordinary skill in theart. See, e.g., Banker and Chalmers, eds., Pharmaceutics and PharmacyPractice, J. B. Lippincott Company, Philadelphia, pp. 238-250 (1982),and Toissel, ASHP Handbook on Injectable Drugs, 4th ed., pp. 622-630(1986). Such solutions can contain anti-oxidants, buffers,bacteriostats, and solutes that render the formulation isotonic with theblood of the intended recipient, and aqueous and non-aqueous sterilesuspensions that can include suspending agents, solubilizers, thickeningagents, stabilizers, and preservatives. The compound or salt of thepresent invention may be administered in a physiologically acceptablediluent in a pharmaceutical carrier, such as a sterile liquid or mixtureof liquids, including water, saline, aqueous dextrose and related sugarsolutions, an alcohol, such as ethanol, isopropanol, or hexadecylalcohol, glycols, such as propylene glycol or polyethylene glycol,dimethylsulfoxide, glycerol ketals, such as2,2-dimethyl-1,3-dioxolane-4-methanol, ethers, such aspoly(ethyleneglycol) 400, an oil, a fatty acid, a fatty acid ester orglyceride, or an acetylated fatty acid glyceride with or without theaddition of a pharmaceutically acceptable surfactant, such as a soap ora detergent, suspending agent, such as pectin, carbomers,methylcellulose, hydroxypropylmethylcellulose, orcarboxymethylcellulose, or emulsifying agents and other pharmaceuticaladjuvants.

Oils useful in parenteral formulations include petroleum, animal,vegetable, or synthetic oils. Specific examples of oils useful in suchformulations include peanut, soybean, sesame, cottonseed, corn, olive,petrolatum, and mineral. Suitable fatty acids for use in parenteralformulations include oleic acid, stearic acid, and isostearic acid.Ethyl oleate and isopropyl myristate are examples of suitable fatty acidesters.

Suitable soaps for use in parenteral formulations include fatty alkalimetal, ammonium, and triethanolamine salts, and suitable detergentsinclude (a) cationic detergents such as, for example, dimethyl dialkylammonium halides, and alkyl pyridinium halides, (b) anionic detergentssuch as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin,ether, and monoglyceride sulfates, and sulfosuccinates, (c) nonionicdetergents such as, for example, fatty amine oxides, fatty acidalkanolamides, and polyoxyethylenepolypropylene copolymers, (d)amphoteric detergents such as, for example, alkyl-beta-aminopropionates,and 2-alkyl-imidazoline quaternary ammonium salts, and (e) mixturesthereof.

The parenteral formulations can contain preservatives and buffers. Inorder to minimize or eliminate irritation at the site of injection, suchcompositions may contain one or more nonionic surfactants having ahydrophile-lipophile balance (HLB) of from about 12 to about 17. Thequantity of surfactant in such formulations will typically range fromabout 5 to about 15% by weight. Suitable surfactants includepolyethylene sorbitan fatty acid esters, such as sorbitan monooleate andthe high molecular weight adducts of ethylene oxide with a hydrophobicbase, formed by the condensation of propylene oxide with propyleneglycol. The parenteral formulations can be presented in unit-dose ormulti-dose sealed containers, such as ampules and vials, and can bestored in a freeze-dried (lyophilized) condition requiring only theaddition of the sterile liquid excipient, for example, water, forinjections, immediately prior to use. Extemporaneous injection solutionsand suspensions can be prepared from sterile powders, granules, andtablets of the kind previously described.

Topical formulations, including those that are useful for transdermaldrug release, are well-known to those of skill in the art and aresuitable in the context of the invention for application to skin.Topically applied compositions are generally in the form of liquids,creams, pastes, lotions and gels. Topical administration includesapplication to the oral mucosa, which includes the oral cavity, oralepithelium, palate, gingival, and the nasal mucosa. In some embodiments,the composition contains at least one active component and a suitablevehicle or carrier. It may also contain other components, such as ananti-irritant. The carrier can be a liquid, solid or semi-solid. Inembodiments, the composition is an aqueous solution. Alternatively, thecomposition can be a dispersion, emulsion, gel, lotion or cream vehiclefor the various components. In one embodiment, the primary vehicle iswater or a biocompatible solvent that is substantially neutral or thathas been rendered substantially neutral. The liquid vehicle can includeother materials, such as buffers, alcohols, glycerin, and mineral oilswith various emulsifiers or dispersing agents as known in the art toobtain the desired pH, consistency and viscosity. It is possible thatthe compositions can be produced as solids, such as powders or granules.The solids can be applied directly or dissolved in water or abiocompatible solvent prior to use to form a solution that issubstantially neutral or that has been rendered substantially neutraland that can then be applied to the target site. In embodiments of theinvention, the vehicle for topical application to the skin can includewater, buffered solutions, various alcohols, glycols such as glycerin,lipid materials such as fatty acids, mineral oils, phosphoglycerides,collagen, gelatin and silicone based materials.

Formulations suitable for oral administration can consist of (a) liquidsolutions, such as a therapeutically effective amount of the inventivecompound dissolved in diluents, such as water, saline, or orange juice,(b) capsules, sachets, tablets, lozenges, and troches, each containing apredetermined amount of the active ingredient, as solids or granules,(c) powders, (d) suspensions in an appropriate liquid, and (e) suitableemulsions. Liquid formulations may include diluents, such as water andalcohols, for example, ethanol, benzyl alcohol, and the polyethylenealcohols, either with or without the addition of a pharmaceuticallyacceptable surfactant, suspending agent, or emulsifying agent. Capsuleforms can be of the ordinary hard- or soft-shelled gelatin typecontaining, for example, surfactants, lubricants, and inert fillers,such as lactose, sucrose, calcium phosphate, and corn starch. Tabletforms can include one or more of lactose, sucrose, mannitol, cornstarch, potato starch, alginic acid, microcrystalline cellulose, acacia,gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium,talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid,and other excipients, colorants, diluents, buffering agents,disintegrating agents, moistening agents, preservatives, flavoringagents, and pharmacologically compatible excipients. Lozenge forms cancomprise the active ingredient in a flavor, usually sucrose and acaciaor tragacanth, as well as pastilles comprising the active ingredient inan inert base, such as gelatin and glycerin, or sucrose and acacia,emulsions, gels, and the like containing, in addition to the activeingredient, such excipients as are known in the art.

The compound or salt of the present invention, alone or in combinationwith other suitable components, can be made into aerosol formulations tobe administered via inhalation. The compounds are preferably supplied infinely divided form along with a surfactant and propellant. Typicalpercentages of active compound are 0.01%-20% by weight, preferably1%-10%. The surfactant must, of course, be nontoxic, and preferablysoluble in the propellant. Representative of such surfactants are theesters or partial esters of fatty acids containing from 6 to 22 carbonatoms, such as caproic, octanoic, lauric, palmitic, stearic, linoleic,linolenic, olesteric and oleic acids with an aliphatic polyhydricalcohol or its cyclic anhydride. Mixed esters, such as mixed or naturalglycerides may be employed. The surfactant may constitute 0.1%-20% byweight of the composition, preferably 0.25%-5%. The balance of thecomposition is ordinarily propellant. A carrier can also be included asdesired, e.g., lecithin for intranasal delivery. These aerosolformulations can be placed into acceptable pressurized propellants, suchas dichlorodifluoromethane, propane, nitrogen, and the like. They alsomay be formulated as pharmaceuticals for non-pressured preparations,such as in a nebulizer or an atomizer. Such spray formulations may beused to spray mucosa.

Additionally, the compound or salt of the present invention may be madeinto suppositories by mixing with a variety of bases, such asemulsifying bases or water-soluble bases. Formulations suitable forvaginal administration may be presented as pessaries, tampons, creams,gels, pastes, foams, or spray formulas containing, in addition to theactive ingredient, such carriers as are known in the art to beappropriate.

It will be appreciated by one of ordinary skill in the art that, inaddition to the aforedescribed pharmaceutical compositions, the compoundor salt of the present invention may be formulated as inclusioncomplexes, such as cyclodextrin inclusion complexes, or liposomes.Liposomes serve to target the compounds to a particular tissue, such aslymphoid tissue or cancerous hepatic cells. Liposomes can also be usedto increase the half-life of the inventive compound. Liposomes useful inthe present invention include emulsions, foams, micelles, insolublemonolayers, liquid crystals, phospholipid dispersions, lamellar layersand the like. In these preparations, the active agent to be delivered isincorporated as part of a liposome, alone or in conjunction with asuitable chemotherapeutic agent. Thus, liposomes filled with a desiredinventive compound or salt thereof, can be directed to the site of aspecific tissue type, hepatic cells, for example, where the liposomesthen deliver the selected compositions. Liposomes for use in theinvention are formed from standard vesicle-forming lipids, whichgenerally include neutral and negatively charged phospholipids and asterol, such as cholesterol. The selection of lipids is generally guidedby consideration of, for example, liposome size and stability of theliposomes in the blood stream. A variety of methods are available forpreparing liposomes, as described in, for example, Szoka et al., Ann.Rev. Biophys. Bioeng., 9, 467 (1980), and U.S. Pat. Nos. 4,235,871,4,501,728, 4,837,028, and 5,019,369. For targeting to the cells of aparticular tissue type, a ligand to be incorporated into the liposomecan include, for example, antibodies or fragments thereof specific forcell surface determinants of the targeted tissue type. A liposomesuspension containing a compound or salt of the present invention may beadministered intravenously, locally, topically, etc. in a dose thatvaries according to the mode of administration, the agent beingdelivered, and the stage of disease being treated.

The invention further provides a method for treating or preventing ahuman galactokinase mediated disease or disorder in a mammal. The methodcomprises administering an effective amount of the compound of formula(I), salt thereof, enantiomers thereof, a mixture of enantiomersthereof, or diastereomers of the invention to a mammal afflictedtherewith. Preferably, the mammal is a human.

The term “mammal” includes, but is not limited to, the order Rodentia,such as mice, and the order Logomorpha, such as rabbits. It is preferredthat the mammals are from the order Carnivora, including Felines (cats)and Canines (dogs). It is more preferred that the mammals are from theorder Artiodactyla, including Bovines (cows) and Swines (pigs) or of theorder Perssodactyla, including Equines (horses). It is most preferredthat the mammals are of the order Primates, Ceboids, or Simioids(monkeys) or of the order Anthropoids (humans and apes). An especiallypreferred mammal is the human. Furthermore, the subject can be theunborn offspring of any of the forgoing hosts, especially mammals (e.g.,humans), in which case any screening of the subject or cells of thesubject, or administration of compounds to the subject or cells of thesubject, can be performed in utero.

The human galactokinase mediated disease or disorder is typically adisease or disorder wherein the production of galactose-1-phosphate isimplicated in the development or progression of the disease or disorder.Patients having a deficiency in a galactose-1-phosphateuridyltransferase (GALT, EC 2.7.7.12) activity can have a potentiallylethal disorder called classic galactosemia. GALT facilitates theconversion of uridine diphosphoglucose (UDP-glucose) andgalactose-1-phosphate (gal-1-p) to uridine diphosphogalactose(UDP-galactose and glucose-1-phosphate. Consequently, GALT deficiencyleads to the unique accumulation of gal-1-p and deficiency ofUDP-galactose and UDP-glucose in patient cells. If untreated, classicgalactosemia can result in severe disease in the newborn period,including liver dysfunction, quickly progressing to liver failure,coagulopathy, coma, and death.

In accordance with an embodiment, the invention provides a method oftreating or preventing classic galactosemia comprising administering toa patient in need thereof a therapeutically effective amount of acompound represented by Formula (I) or a salt thereof.

In accordance with another embodiment, the invention provides a methodof reducing the level of galactose-1-phosphate in a cell comprisingcontacting a cell with a therapeutically effective amount of a compoundrepresented by Formula (I) or (II) or a salt thereof.

In accordance with another embodiment, the invention provides a methodfor treating or preventing a PTEN/AKT mis-regulated cancer in a mammal,comprising administering to the mammal in need of, a therapeutically orprophylactically effective amount of a compound represented by Formula(I) or a salt thereof. It has been reported that the galactose kinase 1(GALK1) gene modifies the PTEN/AKT pathway in a number of human tissuesand human cell lines, and that GALK1 mRNA is over-expressed in colon,head/neck, lung, ovary, pancreas, skin, and stomach tumors. GALK1 iswell expressed in standard tumor cell lines including MDA-MB231T (breasttumor cell line), A549 (non-small cell lung tumor cell line,) U87MG(glioblastoma cell line) and PC-3 (prostate tumor cell line). Thus,inhibition of GALK1 may be useful in treatment of the aforesaid cancers.

“Treating” within the context of the present invention, means analleviation of symptoms associated with a disorder or disease, or haltof further progression or worsening of those symptoms. For example,within the context of treating patients with classic galactosemia,successful treatment may be used in conjunction with dietaryrestrictions aimed at eliminating lactose and galactose in the diet. Inaddition, dietary restrictions are insufficient in treating galactosemiain some patients as the human body can produce galactose endogenouslyfrom the natural turnover of glycolipids and glycoproteins. Within thecontext of treating patients with classic galactosemia, successfultreatment may include a reduction in clinical markers such as levels ofgalactose in blood or urine, and/or changes in clinical symptoms such ashepatomegaly (an enlarged liver), cirrhosis, renal failure, cataracts,brain damage, and ovarian failure, and the like. Treatment may alsoinclude administering the pharmaceutical formulations of the presentinvention in combination with other therapies. For example, thecompounds and pharmaceutical formulations of the present invention maybe administered on a chronic basis. The compounds of the invention canalso be administered in conjunction with dietary restrictions.

“Preventing” within the context of the present invention, refers to aprophylactic treatment of an individual prone or subject to developmentof a condition, in particular, a disease or disorder responsive toinhibition of galactokinase. For example, those of skill in the medicalarts may be able to determine, based on clinical symptoms and patienthistory, a statistical predisposition of a particular individual to thedevelopment of the aforesaid disease or disorder. For example, a familyhistory of galactosemia can be used to assess the predisposition of aparticular individual to the development of galactosemia and relateddisorders and thus inform the individual as to the desirability ofpreventative treatment with a compound or salt of the invention or amedicament formed therefrom. Accordingly, an individual predisposed tothe development of a disease or disorder responsive to inhibition ofgalactokinase may be treated with a compound or a composition of thepresent invention in order to prevent, inhibit, reduce the effect of adevelopment of, or slow the development of the disease or disorder orameliorate the condition.

One skilled in the art will appreciate that suitable methods ofutilizing a compound and administering it to a human for the treatmentor prevention of disease states, in particular, classic galactosemia andcancers, e.g., colon, head/neck, lung, ovary, pancreas, skin, breast,non-small cell lung, glioblastoma, prostate, and stomach cancer, whichwould be useful in the method of the present invention, are available.Although more than one route can be used to administer a particularcompound, a particular route can provide a more immediate and moreeffective reaction than another route. Accordingly, the describedmethods are merely exemplary and are in no way limiting.

The dose administered to a mammal, particularly, a human, in accordancewith the present invention should be sufficient to effect the desiredresponse. Such responses include reversal or prevention of the adverseeffects of the disease for which treatment is desired or to elicit thedesired benefit. One skilled in the art will recognize that dosage willdepend upon a variety of factors, including the age, condition, and bodyweight of the human, as well as the source, particular type of thedisease, and extent of the disease in the human. The size of the dosewill also be determined by the route, timing and frequency ofadministration as well as the existence, nature, and extent of anyadverse side-effects that might accompany the administration of aparticular compound and the desired physiological effect. It will beappreciated by one of skill in the art that various conditions ordisease states may require prolonged treatment involving multipleadministrations.

Suitable doses and dosage regimens can be determined by conventionalrange-finding techniques known to those of ordinary skill in the art.Generally, treatment is initiated with smaller dosages that are lessthan the optimum dose of the compound. Thereafter, the dosage isincreased by small increments until the optimum effect under thecircumstances is reached. The present inventive method typically willinvolve the administration of about 0.1 to about 300 mg of one or moreof the compounds described above per kg body weight of the mammal.

By way of example and not intending to limit the invention, the dose ofthe pharmaceutically active agent(s) described herein for methods ofpreventing diseases or disorders, e.g., classic galactosemia andcancers, can be about 0.001 to about 1 mg/kg body weight of the subjectbeing treated per day, for example, about 0.001 mg, 0.002 mg, 0.005 mg,0.010 mg, 0.015 mg, 0.020 mg, 0.025 mg, 0.050 mg, 0.075 mg, 0.1 mg, 0.15mg, 0.2 mg, 0.25 mg, 0.5 mg, 0.75 mg, or 1 mg/kg body weight per day.The dose of the pharmaceutically active agent(s) described herein formethods of treating diseases or disorders, e.g., classic galactosemiaand cancers, can be about 1 to about 1000 mg/kg body weight of thesubject being treated per day, for example, about 1 mg, 2 mg, 5 mg, 10mg, 15 mg, 0.020 mg, 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250mg, 500 mg, 750 mg, or 1000 mg/kg body weight per day.

The compounds of the invention can be synthesized by any suitablemethod, for example, by reacting a 1,3-dicarbonyl compound, an aldehyde,and a guanidine derivative to form the dihydropyrimidine ring. Forexample, the compounds of the invention can be synthesized according tothe following route:

wherein R¹, R², R³, R⁴, and R⁵ are as defined herein.

All air and/or moisture sensitive reactions were performed underpositive pressure of nitrogen with oven-dried glassware. Anhydroussolvents such as dichloromethane, N,N-dimethylformamide (DMF),acetonitrile, methanol and triethylamine were obtained by purchasingfrom Sigma-Aldrich. Preparative purification was performed on a Waterssemi-preparative HPLC. The column used was a Phenomenex Luna C18 (5micron, 30×75 mm) at a flow rate of 45 ml/min. The mobile phaseconsisted of acetonitrile and water (each containing 0.1%trifluoroacetic acid). A gradient of 10% to 50% acetonitrile over 8minutes was used during the purification. Fraction collection wastriggered by UV detection (220 nM). Analytical analysis was performed onan Agilent LC/MS (Agilent Technologies, Santa Clara, Calif.).

Method 1: A 7 minute gradient of 4% to 100% Acetonitrile (containing0.025% trifluoroacetic acid) in water (containing 0.05% trifluoroaceticacid) was used with an 8 minute run time at a flow rate of 1 mL/min. APhenomenex Luna C18 column (3 micron, 3×75 mm) was used at a temperatureof 50° C.

Method 2: A 3 minute gradient of 4% to 100% Acetonitrile (containing0.025% trifluoroacetic acid) in water (containing 0.05% trifluoroaceticacid) was used with a 4.5 minute run time at a flow rate of 1 mL/min. APhenomenex Gemini Phenyl column (3 micron, 3×100 mm) was used at atemperature of 50° C.

Purity determination was performed using an Agilent Diode Array Detectoron both Method 1 and Method 2. Mass determination was performed using anAgilent 6130 mass spectrometer with electrospray ionization in thepositive mode. ¹H NMR spectra were recorded on Varian 400 MHzspectrometers. Chemical Shifts are reported in ppm withtetramethylsilane (TMS) as internal standard (0 ppm) for CDCl₃ solutionsor undeuterated solvent (DMSO-h6 at 2.49 ppm) for DMSO-d6 solutions. Allof the analogs for assay have purity greater than 95% based on bothanalytical methods. High resolution mass spectrometry was recorded on anAgilent 6210 Time-of-Flight LC/MS system. Confirmation of molecularformula was accomplished using electrospray ionization in the positivemode with the Agilent Masshunter software (version B.02).

The following examples further illustrate the invention but, of course,should not be construed as in any way limiting its scope.

Example 1

This example demonstrates a synthesis of an exemplary compound inaccordance with an embodiment of the invention.

Cyclohexane-1,3-dione (0.191 g, 1.703 mmol, 1.5 equiv.) and1-(benzo[d]oxazol-2-yl)guanidine (0.2 g, 1.135 mmol, 1.0 equiv.) wereadded to a 2-5 ml Biotage microwave vial with a stir bar, and thepowders were mixed well. Cyclopentanone (0.102 ml, 1.135 mmol, 1.5equiv.) was added, then the microwave vial was capped and quicklydropped into an oil bath at 120° C. with vigorous stirring. The vial wasstirred at 120° C. for 6 hours, then removed from the oil bath; thereaction flask was carefully vented using a needle, then the cap wasremoved and ˜6 ml of DMSO was added to the hot flask. Once all of thecompound has dissolved, purification was done by directly injecting to aWaters™ reverse phase purification system to giveNCGC00187642/CID:664331 as a TFA salt (0.072 g, 14%).

¹H NMR (400 MHz, DMSO-d₆); 10.37 (br. s. 1H), 9.98 (br. S. 1H), 7.40 (m,2H), 7.16 (m, 2H), 2.52 (m, 2H), 2.39 (m, 2H), 2.26 (m, 2H), 1.83 (m,6H), 1.62 (m, 2H). Method 1, retention time, 5.671 min; Method 2,retention time 3.704 min; HRMS: m/z (M+H+)=336.1592 (Calculated forC₁₉H₂₀N₄O₂=336.1586). Solubility (PBS, pH 7.4, 23° C.)=2.3 μg/ml.

Example 2

This example demonstrates a primary qHTS assay for inhibitors of GALK.

Assay details and protocol: The primary assay monitored ATP depletionusing Promega's KinaseGlo™ technology, where ATP levels are measuredthrough luminescence generated from firefly luciferase, a bioluminescentATP-dependent enzymes. ATP was held at 35 μM, near its reported K_(M)value, and the K_(M) for galactose was determined under the 1536-wellassay conditions to be 50-100M (FIG. 1A). As well, the IC₅₀ for acommercially available CD45 inhibitor(N-(9,10-dioxo-9,10-dihydrophenanthren-2-yl)pivalamide) was confirmed(previously found to inhibit GALK (FIG. 1B)). This was used as thepositive control for the assay. The assay used 5 nM GALK and a 1 hrincubation time, which gave sufficient signal:background and stabilityfor the HTS. The percent conversion of ATP under these conditions wasestimated to be approximately 50% using an ATP standard curve.

Example 3

As in the primary screen, the confirmatory assay monitored ATP depletionusing Promega's KinaseGlo™ technology, where ATP levels are measuredthrough luminescence generated from firefly luciferase, a bioluminescentATP-dependent enzymes, 6. ATP was held at 35 μM, near its reported K_(M)value, and the K_(M) for galactose was determined under the 1536-wellassay conditions to be 50-100 μM.

Example 4 Secondary Assays

The selectivity for these analogs against CDP-ME kinase, another memberof the GHMP kinase family, was performed using a bioluminescent KinaseGlo™ assay that detects ATP depletion after kinase reaction (PubChem ID2506). NCGC00187642 (CID_664331); all analogs showed no activity in thisassay (up to 57 μM), highlighting the selectivity of this chemotype. Allanalogs were also tested for cytotoxicity using Promega CellTiter Glo onHEK293 cells that were treated with compounds and analyzed after 48hours. Compounds 3, 4, 6, 8, 10, 20, 26, 27, 28, 33, and 35 shown intable 1 showed no cytotoxic effect (PubChem ID 2547). In addition, theability of these compounds to undergo redox recycling, which may lead tofalse positive results, was also evaluated using an endpointcolorimetric assay; this assay detects the presence of H₂O₂ in thekinase reaction buffer (PubChem ID 2502). The lead compound and analogsare inactive in the redox recycling, further confirming that thesecompounds have genuine GALK target activity.

Example 5

This example illustrates some of the properties of inventive compoundsof formula I, in accordance with an embodiment of the invention.

The compounds set forth in Table 1 were screened against GALK using theprimary qHTS assay described in Example 1. In addition, Table 1 presentsthe high resolution mass spectrometry molecular weight data for theinventive compounds.

TABLE 1 GALK Assay High Resolution Mass Spec Data IC50 Calcd. Found No.Structure (μM) Efficacy Exact MW [M + H] [M + H] 3

16.7717 85.3762 322.142976 323.1503 323.1509 4

4.2129 92.8022 336.158626 337.1659 337.1664 5

32.6092 47.1996 310.142976 311.1503 311.1513 6

42.1285 42.7443 426.205576 427.2129 427.2140 7

5.1682 42.6121 378.205576 379.2129 379.2132 8

6.6769 86.616 364.189926 365.1972 365.1981 9

14.9478 40.4611 412.189926 413.1972 413.1988 10

13.3222 79.8676 322.142976 323.1503 323.1515 12

47.269 42.9838 412.189926 413.1972 413.1982 14

11.8734 79.2546 350.174276 351.1816 351.1823 16

9.4314 60.4274 378.088353 379.0956 379.0968 17

0.5951 79.2645 392.104004 393.1113 393.1118 19

21.1143 40.3635 364.189926 365.1972 365.1987 20

6.6769 71.2193 370.119654 371.1269 371.1279 21

2.6581 88.8058 354.115047 355.1223 355.1235 22

1.0582 77.7805 368.130697 369.1380 369.1392 23

33.4639 56.2261 352.153541 353.1608 353.1615 24 thiophene dioxide 2-6.6769 67.583 386.104876 387.1122 387.1122 (benzo[d]oxazol-2-ylamino)-4′,5′,7,8-tetrahydro-1H,2′H- spiro[quinazoline-4,3′-thiophen-1,1-dioxide]-5(6H)-one 25

18.8181 36.5021 412.189926 413.1972 413.1982 26

5.9508 80.9702 350.174276 351.1816 351.1829 27

7.4916 73.6075 414.069138 415.0764 415.0768 28

21.1143 65.7113 350.174276 351.1816 351.1830 29

0 0 364.189926 365.1972 365.1979 33

37.5471 32.1721 386.174276 387.1816 387.1824 34

0 0 425.221561 426.2288 35

74.9163 32.4987 349.190260 350.1975 350.1984 36

8.4057 70.3653 352.153541 353.1608 353.1616 37

61.0781 36.3833 392.104004 393.1113 393.1112 38

68.5308 50.426 392.104004 393.1113 393.1118 39

12.1867 95.0778 426.130360 427.1376 427.1389 40

16.7717 21.752 426.130360 427.1376 427.1382 41

0.7492 90.43 436.053488 437.0608 437.0628 42

68.5308 42.0422 376.133554 377.1408 377.1411 43

48.5161 33.6539 388.153541 389.1608 389.1612 44

18.8181 77.0421 388.153541 389.1608 389.1615 45

61.0781 36.6772 388.153541 389.1608 389.1620 46

61.0781 38.7514 372.158626 373.1659 373.1658 47

61.0781 33.0611 372.158626 373.1659 373.1668 48

61.0781 69.6567 378.115047 379.1223 379.1222 49

76.8928 43.3244 359.138225 360.1455 360.1464 50

42.1285 25.6893 359.138225 360.1455 360.1465 51

61.0781 51.6101 376.133554 377.1408 377.1415 52

34.3468 96.8331 358.142976 359.1503 359.1513 53

76.8928 42.2083 401.185175 402.1925 402.1926 55

15.3421 91.8683 393.099252 394.1065 394.1068 57

2.6581 93.8521 392.104004 393.1113 393.1126 58

6.6769 91.8248 427.125609 428.1329 428.1338 59

6.6769 95.2834 427.125609 428.1329 428.1327 60

12.982 46.3283 426.065031 427.0723 427.0736 61

18.8181 60.4156 393.099252 394.1065 394.1072 63

14.566 44.8906 437.048737 438.0560 438.0562 64

36.5881 30.0153 396.110152 397.1174 397.1179 65

9.1905 98.61 378.115047 379.1223 379.1229 66

1.157 86.1547 484.039619 485.0469 485.0480 67

0.6506 73.7449 437.048737 438.0560 438.0570 68

1.4566 90.2363 437.048737 438.0560 438.0570 69

2.9063 97.082 426.043986 427.0513 427.0519 70

2.9063 94.0642 440.059636 441.0669 441.0675 71

20.575 59.2065 437.048737 438.0560 438.0575 73

14.566 56.9903 348.133474 349.1408 349.1410 74

18.3375 64.1609 362.149124 363.1564 363.1576 75

2.9063 69.6318 382.094501 383.1018 405.0853 [M + 23] 76

3.2609 69.0891 535.041065 536.0483 536.0471 77

5.1682 54.5237 535.041065 536.0483 536.0482 78

4.6062 72.3078 515.095688 516.1030 516.1052 79

4.1053 67.2792 515.095688 516.1030 516.1051 80

3.6588 56.7556 531.090602 532.0979 532.0981 81

3.2609 83.3298 531.090602 532.0979 532.0989 82

18.3375 36.4861 525.080038 526.0873 526.0863 83

10.3119 39.4724 529.111338 530.1186 530.1200 84

5.7988 85.8512 492.054551 493.0618 493.0639 85

4.1053 43.3555 508.031707 509.0390 509.0407 86

3.2609 89.641 509.026956 510.0342 510.0355 87

0 0 426.043986 427.0513 427.0514 89

9.1905 61.8961 507.126988 508.1343 508.1358 90

9.1905 64.6859 501.080038 502.0873 502.0889 91

20.575 61.1653 453.080038 454.0873 454.0878 92

6.5064 66.4427 515.095688 516.1030 516.1040 93

3.2609 91.774 409.132094 410.1394 410.1392 94

6.5064 89.4844 392.159689 393.1670 393.1683 95

25.9024 30.4548 458.152495 459.1598 459.1610 96

0 0 442.175339 443.1826 443.1830 97

8.1911 77.2742 452.111214 453.1185 453.1195 98

9.1905 25.4219 518.104020 519.1113 519.1128 99

0 0 502.126864 503.1341 503.1351 100

3.6588 50.3529 545.114919 546.1222 546.1236 101

18.3375 59.169 425.136701 426.1440 426.1455 102

5.1682 75.2589 469.083619 470.0909 470.0914 105

16.3433 64.736 399.121051 400.1283 400.1294 106

16.3433 69.0182 453.168001 454.1753 454.1762 107

11.5702 28.7995 504.088370 505.0957 505.0973 108

5.1682 66.4502 531.099269 532.1065 532.1070 109

18.3375 54.0161 385.105401 386.1127 386.1139 110

20.575 47.4384 411.121051 412.1283 412.1294 111

20.575 24.5579 339.169525 340.1768 340.1776 112

9.1905 86.5744 452.111214 453.1185 453.1195

All references, including publications, patent applications, andpatents, cited herein are hereby incorporated by reference to the sameextent as if each reference were individually and specifically indicatedto be incorporated by reference and were set forth in its entiretyherein.

The use of the terms “a” and “an” and “the” and similar referents in thecontext of describing the invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. All methodsdescribed herein can be performed in any suitable order unless otherwiseindicated herein or otherwise clearly contradicted by context. The useof any and all examples, or exemplary language (e.g., “such as”)provided herein, is intended merely to better illuminate the inventionand does not pose a limitation on the scope of the invention unlessotherwise claimed. No language in the specification should be construedas indicating any non-claimed element as essential to the practice ofthe invention.

Preferred embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention.Variations of those preferred embodiments may become apparent to thoseof ordinary skill in the art upon reading the foregoing description. Theinventors expect skilled artisans to employ such variations asappropriate, and the inventors intend for the invention to be practicedotherwise than as specifically described herein. Accordingly, thisinvention includes all modifications and equivalents of the subjectmatter recited in the claims appended hereto as permitted by applicablelaw. Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the invention unlessotherwise indicated herein or otherwise clearly contradicted by context.

The invention claimed is:
 1. A compound selected from the groupconsisting of:

or a pharmaceutically acceptable salt thereof.
 2. The compound or saltof claim 1, wherein the compound is selected from the group consistingof:


3. The compound or salt of claim 1, wherein the compound is selectedfrom the group consisting of:


4. The compound or salt of claim 1, wherein the compound is selectedfrom the group consisting of:


5. The compound or salt of claim 1, wherein the compound is selectedfrom the group consisting of:


6. The compound or salt of claim 1, wherein the compound is selectedfrom the group consisting of:


7. The compound or salt of claim 1, wherein the compound is selectedfrom the group consisting of:


8. The compound or salt of claim 1, wherein the compound is selectedfrom the group consisting of:


9. The compound or salt of claim 1, wherein the compound is selectedfrom the group consisting of:


10. The compound or salt of claim 1, wherein the compound is selectedfrom the group consisting of:


11. The compound or salt of claim 1, wherein the compound is:


12. The compound or salt of claim 1, wherein the compound is selectedfrom the group consisting of:


13. The compound or salt of claim 1, wherein the compound is selectedfrom the group consisting of: